臺灣博碩士論文加值系統

幾世紀以來在食品工業中肉桂是重要的香料原料之一。許多的研究中顯示台灣特有種的土肉桂(Cinnamomum osmophloeum Kanhira)，其部分成分與肉桂(Cinnamomum cassai)的皮相類似。土肉桂含有cinnamaldehyde、cinnamic acid、benzoic acid及rutin等主要化合物，其中cinnamaldehyde可應用於保健及生物防治。整體而言土肉桂的用途包括殺菌、抗菌、抗細胞毒素和殺蟲劑等。本研究的目的在探討土肉桂成分之季節變化，從2006年1月到2006年12月採集土肉桂的葉子，置於50℃的烘箱內乾燥，並用研磨機粉碎，各取2g粉末加100mL甲醇進行迴流萃取2.5小時。此外利用CO2超臨界萃取，在低、中、高等不同的溫度及壓力條件下進行萃取。所有樣品皆經過濃縮及定容，再經0.45µm的濾膜過濾後，注入高效液相層析中進行成分分析。結果顯示各成分之季節性差異顯著，其中以 cinnamaldehyde 及 rutin含量最高，而CO2超臨界萃取則以65℃，3500psi 的高溫高壓為適當之萃取條件。

Cinnamon has been used as a spice for centuries and its importance today is based on its continued utilization within the food industry. However, the endemic species of C. osmophloeum Kanehira found in Taiwan is being sought after for many other purposes. This specific species has been the focus of significant research because its constituents are similar to Cinnamomum cassia bark oil. Some of the major compounds found within C. osmophloeum are cinnamaldehyde, cinnamic acid, benzoic acid and rutin. Cinnamaldehyde continues to stand out as the most dominant compound and has been isolated and proven to be responsible for many health and environmental applications. However, the interactions of compounds provide unique characteristics and potential uses for C. osmophloeum, some of which include antifungal, antibacterial, cytotoxic, and larvicidal applications. The purpose of this research is to note the degree of seasonal variation within the cinnamon leaves by focusing on the four aforementioned compounds. C. osmophloeum leaves were collected from January to December 2006 and oven dried at 50oC. The grounded leaves were distilled using 2 g of powdered leaves per 100 mL of methanol for 2 hours. Then the essential oil from the leaves of C. osmophloeum Kanehira was extracted by performing CO2 Supercritical Fluid Extraction (CO2 SFE) for 2 hours at three replications of low, medium, and high temperature and pressure treatments. All solutions were vacuum filtered, concentrated and filtered once more by a millipore filter measuring 0.45 µm. The four major component contents were analyzed by HPLC. The results showed that compounds do vary in concentration on a monthly basis, and cinnamaldehyde and rutin were the lowest and highest in concentration respectively; when compared to the other compounds evaluated. The highest concentration of essential oil produced by CO2 SFE was obtained from extraction conditions at high temperature and high pressure, such as 65oC at 3500 psi.

摘要…………….........................…………………………………………I
ABSTRACT……………....................….............………………………. II
ACKNOWLEDGEMENTS……..……………….…..........................….IV
TABLE OF CONTENTS……............…..…….……....….……….…....VI
LIST OF TABLES……………………..………………………………VII
LIST OF FIGURES………….………………...……………………….IX
1.INTRODUCTION……...…...………..…….……………………….....1
1.1 General goals…………………..………………….……………...1
1.2 Specific objectives………..……………….……………………...1
1.3 Rational of study…..…..….......…………….….…….…………...2
1.4 Cinnamomum osmophloeum Kanehira ……..……......….….……2
1.5 CO2 Supercritical Fluid Extraction…...…...……………………...5
2. LITERATURE REVIEW…………………….......……………………7
2.1 Classification of Cinnamomum osmophloeum Kanehira…..….…..7
2.2 Habitat and plant description…………..……………….........……7
2.3 Biological importance of C. osmophloeum…...…………........…...8
2.3.1 Antimicrobial activity of C. osmophloeum constituent compounds….....……..……………………...……………….9
2.3.2 Antifungal activity of C. osmophloeum constituent compounds .……….………………………………………..10
2.3.3 Antioxidant activity of C. osmophloeum constituent . compounds….....……...………………………......…….…..10
2.3.4 Cytotoxic ability of C. osmophloeum constituent compounds…….....….................................................….......10
2.4 Description of essential oil extraction methods…..…………...........11
2.4.1 Traditional Soxhlet extraction…………..……….…….….....11
2.4.2 Sonication assisted extraction.....…..………………….....….12
2.4.3 Supercritical fluid extraction……..…..……………..…...…..13
2.4.4 Chemical compounds evaluated from C. osmophloeum
Kanehira…….........................…………….………..........….15
3. MATERIAL AND METHODS…...……………….….………...…..….16
3.1 C. osmophloeum Kanehira sample acquisition and
preservation…...........................................................................…...16
3.2 Chemicals and reagents…………………………………………...16
3.3 Preparation of C. osmophloeum Kanehira leaves for HPLC
analysis….....................................................................................….17
3.4 HPLC instrumentation and analytical conditions……..…….18
3.5 Concentrations for compound analysis…...…………………….….19
3.6 Compound analysis and HPLC specifications………………….…20
3.7 Sample acquisition and preparation for CO2 supercritical fluid
extraction (SFE)…..……………..………..……….………………20
3.8 CO2 SFE pre-settings and instrumentation........................…..20
3.9 CO2 SFE treatment parameters……………...…….……………….22
3.10 Extraction and collection of extract …………………..……….....22
3.11 Statistical analysis of data................................................................24
4. RESULTS AND DISCUSSION……….…………..……………..…….25
4.1 Component contents of C. osmophloeum Kanehira by HPLC…….25
4.2 Quantitative results and Calibration curves.........................….........26
4.3 Seasonal variation of component contents of C. osmophloeum
Kanehira..…..................................………….…...…………………26
4.4 Seasonal variation of Rutin within C. osmophloeum leaves.......…..27
4.5 Seasonal variation of Benzoic acid within C. osmophloeum
leaves...……………………………………………………….……29
4.6 Seasonal variation of Cinnamic acid within C. osmophloeum leaves………………………………………………………….....30
4.7 Seasonal variation of Cinnamaldehyde within C. osmophloeum
leaves ................................................................................................31
4.8 CO2 Supercritical Fluid Extraction treatment evaluation ………....32
5. CONCLUSION ….....….………..……………………......…………....35
REFERENCES …….…..….…….……………………....…………..…….36
BIOSKETCH….……........……………………………………......…….....40

Babu, P. S., S. Prabuseenivasan, S. Ignacimuthu. 2007. Cinnamaldehyde-A potential antidiabetic agent. Journal of Phytomedicine, 14: 15-22.
Bernardo-Gill, M. G., J. Grenha, J. Santos, P. Cardoso. 2002. Supercritical fluid extraction and characterization of oil from hazelnut. European Journal of Lipid Science and Technology, 104: 402-409.
Brooks, J. S. and P. Feeny, 2004. Seasonal variation in Daucus corota leaf-surface and leaf-tissue chemical profiles. Journal of Biochemical systematics and ecology, 32:769-782.
Canter, H. P., H. Thomas, E. Ernst. 2005. Bringing medicinal plants into cultivation: opportunities and challenges for biotechnology. Trends in Biotechnology, 23:180-185.
Chang, S. T., P. F. Chen, S. C. Chang. 2001. Antibacterial activity of leaf essential oils and their constituents from Cinnamomum osmophloeum. Journal of Ethnopharmacology, 77: 123-127.
Chau, M. T., Y. T. Tung, S.T. Chang. 2007. Antioxidant activities of ethanolic extracts from the twigs of Cinnamomum osmophloeum. Journal of Bioresource Technology. In press.
Cheng, S. S., J. Y. Liu, Y. R. Hsui, S. T. Chang. 2006. Chemical polymorphism and antifungal activity of essential oils from leaves of different provenances of indigenous cinnamon (Cinnamomum osmophloeum). Bioresource Technology, 97: 306-312.
Dean, J. R., and B. Liu. 2000. Supercritical fluid extraction of Chinese herbal medicines: Investigation of extraction kinetics. Phytochemical Analysis, 11: 1-6.
Eller, F. J., and J. W. King. 2000. Supercritical carbon dioxide extraction of cedarwood oil: a study of extraction parameters and oil characteristics. Phytochemical Analysis, 11: 226-231.

Fang, S. H., Y. K. Rao, Y. M. Tzeng. 2004. Cytotoxic effects of trans-cinnamaldehyde from Cinnamomum osmophloeum leaves on human cancer cell lines. International Journal of Applied Science and Engineering, 2: 136-147.
Fang, S. H., Y. K. Rao, Y. M. Tzeng. 2005. Inhibitory effects of flavonol glycosides from Cinnamomum osmophloeum on inflammatory mediators in LPS/IFN-γ-activated murine macrophages. Bioorganic and Medicinal Chemistry, 13: 2381-2388.
Gomes, P. B., V. G. Mata, A. E. Rodrigues. 2007. Production of rose geranium oil using supercritical fluid extraction. The Journal of Supercritical Fluids, 41: 50-60.
Huang, T. C., H.Y. Fu, C. T. Ho, D. Tan, Y. T. Huang, M. H. Pan. 2007. Induction of apoptosis by cinnamaldehyde from indigenous cinnamon Cinnamomum osmophloeum Kaneh through reactive oxygen species production, glutathione depletion, and caspase activation in human leukemia K562 cells. Journal of Food Chemistry, 103: 434 - 443.
Kim, H. O., S.W. Park, H. D. Park. 2004. Inactivation of Escherichia coli O157:H7 by cinnamic aldehyde purified from Cinnamomum cassia shoot. Food Microbiology, 21: 105-110.
Lang, Q., and C.M. Wai. 2001. Supercritical fluid extraction in herbal and natural product studies-A practical review. Talanta, 53: 771-782.
Li, H., L. Pordesimo, J. Weiss. 2004. High intensity ultrasoundassisted extractionof oil from soybeans. Food Research International, 37: 731-738.
Lin, K. H., S. Y. Yeh, M. Y. Lin, M. C. Shih, K. T. Yang, S. Y. Hwang. 2007. Major chemotypes and antioxidative activity of the leaf essential oils of Cinnamomum osmophloeum Kaneh. From a clonal orchard. Food Chemistry, In press.
Luque de Castro, M. D., and M. T. Tena, 1996. Strategies for supercritical fluid extraction of polar and ionic compounds. Trends in analytical chemistry, 15: 32-37.
Luque de Castro, M. D., and L. E. Garcia-Ayuso, 1998. Soxhlet extraction of solid materials: An outdated technique with a promising innovative future. Analytica Chimica Acta, 369: 1-10.
Maldão-Martins, M., A. Palavra, M. L. Beirão da Costa, and M. G. Bernardo-Gil. 2000. Supercritical CO2 extraction of Thymus zygis L. subsp. sylvestris aroma. The Journal of Supercritical Fluids, 18: 25-34.
Marr, R., and T. Gamse, 2000. Use of supercritical fluids for different processes including new developments-a review. Chemical Engineering and processing, 39: 19-28.
Okamura, N., H. Miki, H. Orii, Y. Masaoka, S. Yamashita, H. Kobayashi, A. Yagi. 1999. Simultaneous high-performance liquid chromatographic determination of puerarin, daidzin, paeo-niflorin, liquiritin, cinnamic acid, cinnamaldehyde and glycyrrhizin in Kampo medicines. Journal of Pharmacy and Biomedical Analysis, 19: 603-612.
Sihvonen, M., E. Jarvenpaa, V. Hietaniemi, and R. Huopalahti. 1999. Advances in supercritical carbon dioxide technologies. Trends in Food Science and Technology, 10: 217-222.
Soares, D., C. G. Pereira, M. A. Meireles, E. M. Saraiva. 2007. Leishmanicidal activity of a supercritical fluid fraction obtained from Tabernaemontana catharinensis. Parasitology International, 56: 135-139.
Stammati, A., P. Bonsi, F. Zucco, R. Moezelaar, H. L. Alakomi, A. von wright. 1999. Toxicity of Selected plant Volatiles in Microbial and Mammalian Short-term Assays. Food and Chemical Toxicology 37: 813-823.
Wang, L., and C. L. Weller 2006. Recent advances in extraction of nutraceuticals from plants. Trends in Food Science & Technology, 17: 300-312.
Wang, S. Y., P. F. Chen, S. T. Chang. 2005. Antifungal activities of essential oils and their constituents from indigenous cinnamon (Cinnamomum osmophloeum) leaves against wood decay fungi. Bioresource Technology, 96: 813-818.
Web References:
Berger, T., and D. Hurren. 1999. Supercritical Fluid Extraction with CO2. Retrieved May 24, 2007, from the World Wide Web: http://www.messer.co.uk
Encyclopedia of Spices 2007. Cinnamon Retrieved June 5, 2006, from the World Wide Web: http://www.theepicentre.com/Spices/cinnamon.html
FAO Document repository 1995 Flavors and Fragrances of plant origins, Non-wood forestry products. Retrieved June 6, 2006, from the World Wide Web: http://www.fao.org/documents/show_cdr.asp?url_file=/docrep/V5350E/V5350e03.htm
Pan, F.J. 1998. Cinnamomum osmophloeum. In: IUCN 2006. 2006 IUCN Red List of Threatened Species. Retrieved May 20, 2007 from the world wide web: www.iucnredlist.org
Pollution Prevention Resource Center (PPRC). 2000. Supercritical Carbon-dioxide Cleaning Technology Review. Retrieved June 30, 2007 from the world wide web:http://www.pprc.org/pubs/techreviews/co2/co2intro.html